Dear All, I have been having a problem staining PBMC with CFSE for cell division tracking. Whenever I label PBMC, I seem to get a second population that is dimmer than the rest of the cells. They do not seem to be cells that have divided, as I see this even after an overnight incubation with no overt stimulation. The cells are not T cells and have a FSC/90LS profile matching that of lymphocytes. Often, these dim cells are as much as 20% of undivided cells in the lymphocyte FSC/90LS gate and they persist without apparent division for as long as I have tested: one week. I have seen this with two batches of CFSE, with a range of concentrations and labelling times of CFSE and with PBMC from a variety of donors. Is anyone familiar with what causes this/what these cells are and more importantly, what conditions to use to prevent this? I have never seen such a thing when labelling a large variety of mouse cell types with CFSE. Thank you for any help you can render. Jeff Dorfman ------ Jeffrey Dorfman, Ph.D. Kenya Medical Research Institute Wellcome Trust Research Laboratories Post Office Box 230 Kilifi, Kenya jdorfman@kilifi.mimcom.net jeffreydorfman@yahoo.com Phone +254 125 22063 Fax +254 125 22390 Mobile+254 733 86.23.10 Home +254 125 22053 UK mobile +44 7814 574-997
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